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- Table of Contents
SORT1 / Sortilin · Western blot design guide
Design a Western Blot for SORT1
Real validated SORT1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SORT1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
SORT1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~92.1 kDa | |
| Observed band | ~100 kDa | |
| Gel | 10% (catalog M01666-1) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SORT1 Western Blot Protocols
The M01666-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human HepG2, human U251 (catalog M01666-1) |
| Gel % | 10% (catalog M01666-1) |
| Load | 30 ug; reducing conditions (catalog M01666-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01666-1) |
| Membrane | nitrocellulose membrane (catalog M01666-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01666-1) |
| Primary antibody | M01666-1 · 1:500 (catalog M01666-1) |
| Primary incubation | overnight at 4°C (catalog M01666-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M01666-1) |
| Secondary incubation | 1.5 hour at RT (catalog M01666-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01666-1) |
| Detection | ECL (catalog M01666-1) |
Section 2
What Is the Expected SORT1 Western Blot Band Size?
SORT1 has a predicted precursor mass of 92.1 kDa and an empirical band near 100 kDa; the cause of the difference is not established.
| Band near 100 kDa | Empirical SORT1 band in reducing whole-cell lysates; identity still requires controls |
| Band below the precursor position | Could reflect signal-peptide or propeptide cleavage; its migration is not established |
| Higher band or diffuse signal | Could reflect heterogeneous N-linked glycosylation; the sites alone do not establish a shift or smear |
| Several distinct bands | Could reflect isoforms 1 and 2 or processing; distinct isoform migration is unverified |
| Faint band in a soluble fraction | SORT1 is a membrane protein and may be depleted from that fraction |
💡Expected SORT1 appearanceUniProt predicts a 92.1 kDa precursor, while antibody QC detects SORT1 near 100 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm band identity with controls.
| UniProt predicted precursor mass | 92.1 kDa sequence-based reference, not a measured migration position |
| N-linked glycosylation sites at Asn98, Asn162, Asn274, Asn406, Asn582 and Asn684 | May alter apparent migration, but no shift magnitude is established |
| Signal peptide at residues 1–33 | Cleavage makes the mature chain shorter than the precursor |
| Propeptide at residues 34–77 | Cleavage further shortens the mature chain; its band position is unspecified |
| Splice isoforms 1 and 2 | May differ in size, but their individual masses and band separation are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated SORT1 may be poorly recovered during extraction | Check membrane extraction and use a positive-control whole-cell lysate |
| Band higher than expected | N-linked glycosylation may affect migration; the cause of the observed 100 kDa band is unestablished | Compare with a validated positive control and assess band identity |
| Band lower than expected | Signal-peptide or propeptide cleavage may shorten SORT1 | Check antibody epitope and compare with a validated positive control |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible but unproven | Compare with a positive control and assess glycosylation if needed |
| Multiple bands | Isoforms 1 and 2 or processing are possible; distinct bands are unverified | Check antibody specificity and compare samples with a positive control |
| Weak or no signal | Poor recovery of membrane-associated SORT1 | Check extraction and loading with a positive-control lysate |
Experiment evidence
Western blot images showing SORT1 band
M01666-1 · SORT1
A01666 · SORT1
A01666 · SORT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SORT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SORT1 in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: SORT1 is a membrane protein, so membrane-enriched lysate may improve detection.
HPA tissue expression evidence for SORT1
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | basal cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SORT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SORT1, answered from its protein features.
What should be checked when unexpected SORT1 bands appear?
Band shift · Check antibody epitope coverage against isoform 2’s missing canonical residues 1–136 and the signal peptide and propeptide at 1–33 and 34–77. Also consider the six listed N-linked sites when assessing apparent mass. These features suggest checks to make; they do not identify an unexpected band by themselves.
How could SORT1 isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. Relative to the canonical sequence, isoform 2 lacks residues 1–136 and has KA replaced by T at positions 278–279. Check which isoform the antibody can recognize before assigning a second band; the listed variants do not establish where either isoform will migrate.
Which glycosylation sites matter when assessing SORT1 bands?
PTM · The six listed N-linked sites are asparagines 98, 162, 274, 406, 582, and 684 in canonical UniProt numbering. Isoform 2 lacks canonical residues 1–136, including site 98. Consider isoform identity when comparing bands, but do not infer a specific shift from the site list alone.
UniProt lists phosphoserines at canonical positions 814, 819, and 825. Check whether the antibody targets one of these residues and whether its numbering follows the canonical sequence. The listed sites alone do not predict a detectable mobility shift.
UniProt lists phosphoserines at canonical positions 814, 819, and 825. Check whether the antibody targets one of these residues and whether its numbering follows the canonical sequence. The listed sites alone do not predict a detectable mobility shift.
Does this guide establish induction of SORT1?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for SORT1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01666-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should SORT1 bands be quantified across samples?
Quantitation · Define the band being measured and use the same assignment across samples. SORT1 has two isoforms, cleavable N-terminal regions, and six listed N-linked sites, so a band assignment should account for which forms the antibody recognizes. The features alone do not justify combining every detected band into one measurement.
Why might SORT1 appear near 100 kDa instead of 92.1 kDa?
Interpretation · 92.1 kDa is the predicted mass; the supplied Western blot observation is approximately 100 kDa. SORT1 has six N-linked glycosylation sites and cleavable signal and propeptides. These features are relevant when assessing apparent mass, but their presence alone does not establish the cause of the difference.
The canonical sequence has a signal peptide at residues 1–33 and a propeptide at 34–77. Check whether the antibody epitope lies in either region before using it to detect processed SORT1. Use canonical UniProt coordinates when comparing epitope maps.
The canonical sequence has a signal peptide at residues 1–33 and a propeptide at 34–77. Check whether the antibody epitope lies in either region before using it to detect processed SORT1. Use canonical UniProt coordinates when comparing epitope maps.
Boster reagents
SORT1 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Sortilin Rabbit Monoclonal Antibody
Cat # M01666-1
Real WB data
Anti-Sortilin SORT1 Antibody
Cat # A01666
Two the supplier anti-SORT1 antibodies have Western blot images. M01666-1 shows a band near 100 kDa in four human cell lines, versus an expected 92 kDa. A01666 has a blot caption, but its tested cell lines and species are unnamed; no publication evidence is supplied.
Which to pick: For human cell lysates, M01666-1 has documented K562, HepG2, U251 and MCF-7 examples. A01666 lists human, mouse and rat reactivity, but its blot caption does not identify the tested cell lines or species. Choose based on your sample species and the reported conditions.
Source: BosterBio SORT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99523.
- Human Protein Atlas. SORT1 tissue expression.
- PMC12334890 — target-verified WB comparison
- PMC3929086 — target-verified WB comparison